Acetate → 4-Nitrophenol
反応式
C([O-])(=O)C.C(C(C(C([O-])=O)O)O)([O-])=O.P([O:19][C:20]1[CH:25]=[CH:24][C:23]([N+:26]([O-:28])=[O:27])=[CH:22][CH:21]=1)(O)(O)=O>>[N+:26]([C:23]1[CH:24]=[CH:25][C:20]([OH:19])=[CH:21][CH:22]=1)([O-:28])=[O:27]参加物
4
実験手順
The scales treated by fixation were moved and weighed. After measurement, the scales were put into a 96-well microplate. To each well, 2001 of 100 mM acetate buffer (pH 5.3) containing 20 mM tartrate and 10 mM para-nitrophenol phosphate (substrate) was added. Reaction was performed at 25° C. for 20 minutes and subsequently terminated by the addition of a 3 M sodium hydroxide aqueous solution (50±1). Then, 1501 of the solution after the completion of reaction was transferred to another microplate. The amount of para-nitrophenol (p-NP) produced by TRAP was measured with a spectrophotometer (405 nm). The osteoclast activity was indicated as the amount of p-NP produced by the decomposition of para-nitrophenol phosphate per g of scale per minute.